[Automated-sequencing] variation in libraries

Fulton, Ciaran ciaran.Fulton at arradx-almac.com
Tue Sep 20 08:55:33 EST 2005


Hi...I was wondering if anyone has had experience of variable length of read statistics within and between different 96-well plates for specific cDNA libraries. We used to pick manually into 96 DWP but increasing throughput meant we moved over to a QPix. The results haven't been great with it but we need to optimise it. But when we went back to picking by hand the results were awful. We normally generate the cDNA libraries using the Invitrogen CloneMiner kit and electroporate into DH10B cells. However, another transformation attempt at the same library by heat shock and into DH5-alpha cells has meant a massive drop in yield across the plate. The LOR results are so variable between plates that we are wrecking our heads trying to come up with answers. Done al the usual yield increasing steps we can think of...Terrific broth, increasing time etc, but no luck. Any suggestions

Dr. Ciaran Fulton
Senior Scientist

ArraDx Ltd
19 Seagoe Industrial Estate
Craigavon 
Co Armagh
BT63 5QD
Northern Ireland




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